大蚕、蛹组织切片核酸特异染色技术的探讨

本文对大蚕、蛹的石蜡切片制作及其核酸特异染色方法进行探讨，针对虫体大和蛹体壁高度丁质化特点，对组织固定，脱水、透明、透蜡的时间相应的增加和固定前蛹皮剪开，在截位时要考虑切片段两头保留一段仅用来起保护作用的部分，并采用了二种核酸特异染色方法染色。结果表明：制片设计方案基本可行，部分细节需进一步改进；甲基绿-派罗宁（昂纳-帕彭海姆）染色法的切片组织颜色层次分明，DNA含量丰富的丝腺组织呈紫黑色，其它组织的DNA呈紫红色，RNA呈淡红色，改良的甲基绿一派罗宁染色法的切片组织颜色偏淡；孚尔根染色法的切片组织颜色层次分明，核酸呈紫红色，其它无色，能较好的以颜色的深浅来判断核酸的丰度。更理想的染色方案还有待进一步摸索。     

Serodiagnosis of Burkholderia mallei Infections in Horses: State‐of‐the‐art and Perspectives

Burkholderia mallei causes glanders or farcy in solipeds, a disease that must be reported to the OIE (Office International des Epizooties, Paris, France). The number of reported outbreaks has increased steadily during the last decade. Serodiagnosis is hampered by the considerable number of false‐positives and ‐negatives of the internationally prescribed tests. The major problem leading to low sensitivity and specificity of complement fixation test (CFT) and enzyme‐linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e. crude preparations of whole cells. Future perspectives for the development and evaluation of serological test kits using well‐characterized single antigens are discussed in the light of recent molecular research on B. mallei and the closely related saprozoonotic agent B. pseudomallei.     

Short-distance wind dispersal of the fungal pathogens causing Sigatoka diseases in banana and plantain

Airborne spores of the fungal pathogens causing Sigatoka diseases in banana and plantain were monitored using rotorod spore traps, sited at various heights within an infected plantation in Costa Rica from December 1993 to February 1994. Different capture patterns of ascospores and conidia were found and the relationship between wind behaviour and spore catches was investigated. This information has enabled an assessment to be made of the reliability of point measurements of airborne spores for monitoring spore movements on the plantation scale. The use of such information in forecasting the airborne movement of these spores and the likely role of the wind in the spread of this disease to uninfected areas is discussed.     

Transfer from Erwinia herbicola to Escherichia coli of a plasmid associated with biocontrol of fire blight

Strains of Erwinia herbicola effective in the biocontrol of fire blight of hawthorn were used to investigate the possibility that the antagonistic activity is coded by plasmid-born genes. Agarose gel electrophoresis of isolated plasmids from four antagonistic Erw. herbicola strains showed a band of a supercoiled 12 kb plasmid in each strain, with a second band greater than 16.2 kb consistently seen in two strains. Erw. herbicola strains showed resistance to penicillin-G, which could be conferred on penicillin-G sensitive Escherichia coli TG1 by transformation with a pure Erw. herbicola plasmid preparation. Transformed strains of Esc. coli appeared to contain the Erw. herbicola 12 kb plasmid, but not the > 16.2 kb plasmid. In an agar plate assay, Esc. coli transformants produced an inhibition zone against Erw. amylovora similar to those produced by the original Erw. herbicola strains. In two biocontrol assays, the transformed Esc. coli strains had a suppressive effect on disease development on infected pear fruit slices and hawthorn blossoms.     

Beschreibung des Feuerbrandbefalls am Institut für Obstzüchtung in Dresden-Pillnitz im Jahr 2003

The orchard of the Institute of Fruit Breeding of the German Federal Centre of Breeding Research on Cultivated Plants in Dresden-Pillnitz was highly affected by fire blight in 2003. Infected pomefruit trees were observed over a period of nearly 3 months. The first symptoms on pear trees were found on May 19th. The pathogen Erwinia amylovora was confirmed officially on May 26, and the last infected apple trees were detected the 11th of August. The infected trees had to be grubbed at the decision of the Phytopathological Authority. In total, 1164 apple and 478 pear trees were grubbed, including the entire pear collection of the gene bank. Of 35 wild species of pear, 49 accessions, eight accessions of six species each, showed infections. The apple collection of the gene bank included 33 wild species, with 365 accessions, and 845 cultivars and clones. Ten accessions of nine wild apple species and 81 cultivars/clones of these collections showed fire blight infection. The source of infection was the pear collection, and the distance from that source was important for the occurrence of infection. Field plots close to the pear collection had tree losses of 10–34%, while more distant plots had losses of 0–6%. Around 80% of the lost apple trees were detected and grubbed from 27th May to 11th June. Some of the cultivars bred in Dresden-Pillnitz, e.g. ‘Pilot’ and ‘Rekarda’, were affected by fire blight in most field plots, whereas most others were affected mainly only in plots adjacent to the infection source. A correlation of r=?0.72 could be calculated for rating in artificial shoot inoculations and percentage of trees of resistant cultivars lost. The cultivars ‘Pirol’, ‘Reanda’, ‘Remo’, ‘Rene’, ‘Renora’, ‘Resi’, and ‘Retina’ showed only a very low numbers of infected trees. No tree of ‘Rewena’ showed symptoms of fire blight. Despite a tendency to postblooming, only 8.9% of ‘Pinova’ trees had to be grubbed.     

Comparison of a commercial H1N1 enzyme-linked immunosorbent assay and hemagglutination inhibition test in detecting serum antibody against swine influenza viruses.

Recently a commercial enzyme-linked immunosorbent assay (ELISA) kit for detecting antibody against H1N1 swine influenza virus (SIV) has been made available to diagnosticians and veterinary practitioners. Because the hemagglutination inhibition (HI) test has been considered the standard test for SIV serology, diagnostic performance of the new ELISA was evaluated using positive (n = 60) and negative (n = 188) serum samples from young pigs with known status of SIV infection and compared with that of the HI test. Both ELISA and HI test identified all negative animals correctly. None of the serum samples (n = 64) from pigs inoculated with H3N2 SIV was positive by ELISA for SIV antibody. The H1N1 SIV antibody detectable by ELISA appears to develop more slowly in comparison with antibody detectable by HI test. Although antibody was detected by HI test in all inoculated animals (n = 20) by day 7 postinoculation (PI), antibody was detected by ELISA in 0%, 75%, and 100% of the inoculated animals on days 7, 14, and 28 PI, respectively. Discrepancy in test results between the 2 serologic tests appeared to be because of differences in antibody isotypes detected by each test. Enzyme-linked immunosorbent assay mainly detected IgG antibody, whereas the HI test detects IgM antibody very efficiently as well as IgG antibody. Collectively, the commercial ELISA is highly specific for antibody to H1N1 SIV but may not identify positive animals at the early stage of infection as effectively as the HI test, particularly when SIV is introduced to a na?ve swine population.